Journal: Molecular Cancer

Article Title: HnRPD/AUF1 facilitates human ovarian cancer progression through activating FLI1 and maintaining cisplatin resistance

doi: 10.1186/s12943-026-02599-5

Figure Lengend Snippet: HnRPD is identified to be an RAB8A mRNA-interacting protein. A The mRNA levels of RAB8A in OVCAR-8 cells transfected with a vector expressing GPR137 or a control empty vector (Con) and cultured with actinomycin D (AcD) for the indicated time periods. * p < 0.05; ** p < 0.01 versus Con; error bar, SD. N = 3. B qRT-PCR analysis of co-precipitated RAB8A mRNA by GPR137 antibody in the RIP assay in SK-OV-3 cells. IgG was used as a negative control. N = 3. C qRT-PCR analysis of co-precipitated RAB8A mRNA by GPR137 antibody in the RIP assay in A2780 cells. IgG was used as a negative control. N = 3. D An experimental model showing the procedure of IP and RNA pull-down with LC-MS/MS analysis. E LC-MS/MS analysis showing the identified proteins with difference in abundance from GPR137 antibody-IP group versus the corresponding control. F LC-MS/MS analysis showing the identified proteins with difference in abundance from RAB8A RNA pull-down group versus the corresponding control

Article Snippet: GPR137 (bs-16270R, 1:1000) antibody and RAB8A (bs-6176R, 1:1000) antibody were acquired from Bioss Biotechnology (Beijing, China), while Flag-tag (M185-3, 1:10000) antibody was obtained from MBL Biotechnology (Beijing, China).

Techniques: Transfection, Plasmid Preparation, Expressing, Control, Cell Culture, Quantitative RT-PCR, Negative Control, Liquid Chromatography with Mass Spectroscopy

Journal: Molecular Cancer

Article Title: HnRPD/AUF1 facilitates human ovarian cancer progression through activating FLI1 and maintaining cisplatin resistance

doi: 10.1186/s12943-026-02599-5

Figure Lengend Snippet: Molecular docking analysis of GPR137-hnRPD and hnRPD- RAB8A RNA. A Docking results by HDOCK server. Yellow indicates GPR137, purple indicates hnRPD. B Docking results visualized in the form of Surface. Red indicates protein-protein contact area. C Interaction of GPR137-hnRPD analyzed by PLIP. D Docking results by HDOCK server. Green indicates RAB8A RNA, purple indicates hnRPD. E Docking results visualized in the form of Surface. Red indicates protein-RNA contact area. F Interaction of hnRPD-RAB8A RNA analyzed by PLIP

Article Snippet: GPR137 (bs-16270R, 1:1000) antibody and RAB8A (bs-6176R, 1:1000) antibody were acquired from Bioss Biotechnology (Beijing, China), while Flag-tag (M185-3, 1:10000) antibody was obtained from MBL Biotechnology (Beijing, China).

Techniques:

Journal: Molecular Cancer

Article Title: HnRPD/AUF1 facilitates human ovarian cancer progression through activating FLI1 and maintaining cisplatin resistance

doi: 10.1186/s12943-026-02599-5

Figure Lengend Snippet: GPR137 promotes RAB8A mRNA stability through elevating hnRPD expression. A Co-IP of endogenous GPR137 and hnRPD in SK-OV-3 cells. IP: GPR137, WB: hnRPD. IgG was used as a negative control. B Co-IP of endogenous hnRPD and GPR137 in A2780 cells. IP: hnRPD, WB: GPR137. IgG was used as a negative control. C qRT-PCR analysis of co-precipitated RAB8A mRNA by hnRPD antibody in the RIP assay in SK-OV-3 cells. IgG was used as a negative control. ** p < 0.01 versus IgG; error bar, SD. N = 3. D RNA pull-down assay in A2780 cells with RAB8A RNA or control RNA fragments. Protein abundance was normalized to GAPDH. E The mRNA expression of hnRPD in SK-OV-3 cells transfected with GPR137 shRNA or scrambled shRNA (shRNA-Con). Error bar, SD. N = 3. F The mRNA expression of hnRPD in OVCAR-8 cells transfected with a vector expressing GPR137 or a control empty vector (Con). Error bar, SD. N = 3. G The protein expression of GPR137 and hnRPD in SK-OV-3 cells transfected with GPR137 shRNA (shGPR137, +) or scrambled shRNA (shGPR137, -) and cultured for 48 h. H The protein expression of GPR137 and hnRPD in A2780 cells transfected with GPR137 shRNA (shGPR137, +) or scrambled shRNA (shGPR137, -). I The protein expression of Flag and hnRPD in OVCAR-8 cells transfected with a vector expressing Flag-GPR137 (Flag-GPR137, +) or a control Flag-tagged empty vector (Flag-GPR137, -). J Western blot analysis for GPR137 and hnRPD in SK-OV-3 cells transfected with GPR137 shRNA (shGPR137, +) or scrambled shRNA (shGPR137, -) and treated for different time periods with CHX. K Western blot analysis for Flag, GPR137 and hnRPD in OVCAR-8 cells transfected with a vector expressing Flag-GPR137 (Flag-GPR137, +) or a control Flag-tagged empty vector (Flag-GPR137, -) and treated for different time periods with CHX. L Western blot analysis for hnRPD in SK-OV-3 cells transfected with GPR137 shRNA (shGPR137, +) or scrambled shRNA (shGPR137, -) and treated with or without MG132. M The mRNA expression of RAB8A in SK-OV-3 cells transfected with hnRPD shRNA or scrambled shRNA (shRNA-Con). ** p < 0.01 versus shRNA-Con; error bar, SD. N = 3. N The mRNA expression of RAB8A in OVCAR-8 cells transfected with a vector expressing hnRPD or a control empty vector (Con). ** p < 0.01 versus Con; error bar, SD. N = 3. O The mRNA levels of RAB8A in SK-OV-3 cells transfected with hnRPD shRNA (sh hnRPD) or scrambled shRNA (shCon) and cultured with actinomycin D (AcD) for the indicated time periods. * p < 0.05; ** p < 0.01 versus shCon; error bar, SD. N = 3. P The mRNA levels of RAB8A in OVCAR-8 cells transfected with a vector expressing hnRPD or a control empty vector (Con) and cultured with actinomycin D (AcD) for the indicated time periods. * p < 0.05; ** p < 0.01 versus Con; error bar, SD. N = 3. Q The mRNA levels of RAB8A in OVCAR-8 cells transfected with a vector expressing GPR137 or a control empty vector (Con) in combination with hnRPD shRNA (sh hnRPD) or scrambled shRNA and cultured with actinomycin D (AcD) for the indicated time periods. * p < 0.05; ** p < 0.01 versus Con; # p < 0.05 versus GPR137; error bar, SD. N = 3. R GEPIA database displayed the correlation between RAB8A and hnRPD

Article Snippet: GPR137 (bs-16270R, 1:1000) antibody and RAB8A (bs-6176R, 1:1000) antibody were acquired from Bioss Biotechnology (Beijing, China), while Flag-tag (M185-3, 1:10000) antibody was obtained from MBL Biotechnology (Beijing, China).

Techniques: Expressing, Co-Immunoprecipitation Assay, Negative Control, Quantitative RT-PCR, Pull Down Assay, Control, Quantitative Proteomics, Transfection, shRNA, Plasmid Preparation, Cell Culture, Western Blot

Journal: Molecular Cancer

Article Title: HnRPD/AUF1 facilitates human ovarian cancer progression through activating FLI1 and maintaining cisplatin resistance

doi: 10.1186/s12943-026-02599-5

Figure Lengend Snippet: GPR137-hnRPD-RAB8A axis regulates OC cell biological behaviors. A CCK-8 assays of OVCAR-8 cells transfected with a GPR137-expressing vector or an empty vector (in Control group) in combination with hnRPD shRNA (shhnRPD) or scrambled shRNA (in Control group) and cultured for the indicated time periods. ** p < 0.01 versus Control, ## p < 0.01 versus GPR137; error bar, SD. N = 3. B Wound healing assays of OVCAR-8 cells transfected with a GPR137-expressing vector or an empty vector (in Control group) in combination with hnRPD shRNA (shhnRPD) or scrambled shRNA (in Control group) at 24 h. Migrated distance was statistically analyzed. Bar, 100 μm. ** p < 0.01 versus shRNA-Con+Control, ## p < 0.01 versus shRNA-Con+GPR137; error bar, SD. N = 3. C CCK-8 assays of OVCAR-8 cells transfected with a hnRPD-expressing vector or an empty vector (in Control group) in combination with RAB8A shRNA (shRAB8A) or scrambled shRNA (in Control group) and cultured for the indicated time periods. * p < 0.05 versus Control, # p < 0.05 versus hnRPD, ** p < 0.01 versus Control, ## p < 0.01 versus hnRPD; error bar, SD. N = 3. D Wound healing assays of OVCAR-8 cells transfected with a hnRPD-expressing vector or an empty vector (in Control group) in combination with RAB8A shRNA (shRAB8A) or scrambled shRNA (in Control group) at 24 h. Migrated distance was statistically analyzed. Bar, 100 μm. ** p < 0.01 versus shRNA-Con+Control, ## p < 0.01 versus shRNA-Con+hnRPD; error bar, SD. N = 3. E - H Spearman correlation analysis plots showing the correlation between the pathway scores and the expression of gene hnRPD . In the plots, the x-axis represents the distribution of the expression of hnRPD , and the y-axis represents the distribution of the corresponding pathway score. The values at the top represent the results of the Spearman correlation analysis, including the p -value, and correlation coefficient. I Matrigel invasion assays of OVCAR-8 cells transfected with a GPR137-expressing vector or an empty vector (in Control group) in combination with hnRPD shRNA (shhnRPD) or scrambled shRNA (in Control group) at 24 h. Bar, 100 μm. J Colony formation assays of OVCAR-8 cells infected with lentiviruses expressing GPR137 or control lentiviruses (in Control group) in combination with lentiviruses carrying hnRPD shRNA or scrambled shRNA (in Control group). K Quantitative analysis of (E). ** p < 0.01 versus shRNA-Con+Control, ## p < 0.01 versus shRNA-Con+GPR137; error bar, SD. N = 3. L Quantitative analysis of (F). ** p < 0.01 versus shRNA-Con+Control, ## p < 0.01 versus shRNA-Con+GPR137; error bar, SD. N = 3. M Matrigel invasion assays of OVCAR-8 cells transfected with a hnRPD-expressing vector or an empty vector (in Control group) in combination with RAB8A shRNA (shRAB8A) or scrambled shRNA (in Control group) at 24 h. Bar, 100 μm. N Colony formation assays of OVCAR-8 cells infected with lentiviruses expressing hnRPD or control lentiviruses (in Control group) in combination with lentiviruses carrying RAB8A shRNA or scrambled shRNA (in Control group). O Quantitative analysis of (I). ** p < 0.01 versus shRNA-Con+Control, ## p < 0.01 versus shRNA-Con+hnRPD; error bar, SD. N = 3. P Quantitative analysis of (J). ** p < 0.01 versus shRNA-Con+Control, ## p < 0.01 versus shRNA-Con+hnRPD; error bar, SD. N = 3. Q CCK-8 assays of SK-OV-3 cells transfected with GPR137 shRNA (shGPR137) or scrambled shRNA in combination with a hnRPD-expressing vector or an empty vector (Control) and cultured for the indicated time periods. * p < 0.05 versus Control, # p < 0.05 versus shGPR137; ** p < 0.01 versus Control, ## p < 0.01 versus shGPR137; error bar, SD. R Migrated distance was statistically analyzed in SK-OV-3 cells transfected with GPR137 shRNA (shGPR137) or scrambled shRNA in combination with a hnRPD-expressing vector or an empty vector (Control) at 24 h. ** p < 0.01 versus shRNA-Con+Control, ## p < 0.01 versus shRNA-GPR137 + Control; error bar, SD. S Matrigel invasion assays of SK-OV-3 cells transfected with GPR137 shRNA (shGPR137) or scrambled shRNA (in Control group) in combination with a hnRPD-expressing vector or an empty vector (in Control group) at 24 h. Bar, 100 μm. T Colony formation assays of SK-OV-3 cells infected with lentiviruses carrying GPR137 shRNA or scrambled shRNA (in Control group) in combination with lentiviruses expressing hnRPD or control lentiviruses (in Control group). U Quantitative analysis of (O). ** p < 0.01 versus shRNA-Con+Control, ## p < 0.01 versus shRNA-GPR137 + Control; error bar, SD. N = 3. V Quantitative analysis of (P). * p < 0.05 versus shRNA-Con+Control, # p < 0.05 versus shRNA-GPR137 + Control; error bar, SD. N = 3

Article Snippet: GPR137 (bs-16270R, 1:1000) antibody and RAB8A (bs-6176R, 1:1000) antibody were acquired from Bioss Biotechnology (Beijing, China), while Flag-tag (M185-3, 1:10000) antibody was obtained from MBL Biotechnology (Beijing, China).

Techniques: CCK-8 Assay, Transfection, Expressing, Plasmid Preparation, Control, shRNA, Cell Culture, Infection

Journal: Molecular Cancer

Article Title: HnRPD/AUF1 facilitates human ovarian cancer progression through activating FLI1 and maintaining cisplatin resistance

doi: 10.1186/s12943-026-02599-5

Figure Lengend Snippet: Suppression of hnRPD expression inhibits OC progression in vivo. A Nude mice with xenografts derived from OVCAR-8 cells stably expressing hnRPD or control. N = 6. B Image showing the size of the tumor xenografts from the two groups in (A). C Weights of xenografts in (B). N = 6. ** p < 0.01 versus Con; error bar, SD. D Tumor volumes of xenografts in (B). N = 6. ** p < 0.01 versus Con; error bar, SD. E The mRNA levels of RAB8A in xenografts in (B). N = 6. ** p < 0.01 versus Con; error bar, SD. F RAB8A staining of RAB8A in xenografts in (B). N = 6. Bar, 20 μm. G IHC score for RAB8A staining in (F). ** p < 0.01 versus Con. H Graphic illustration of the intraperitoneal injection of control or hnRPD stable-overexpressing OVCAR-8 cells. I The metastasis and spread of control or hnRPD stable-overexpressing OVCAR-8 cells were monitored by IVIS Imaging System. N = 5. J Nude mice with xenografts derived from OVCAR-8 cells stably expressing GPR137 or in combination with hnRPD shRNA (shhnRPD). N = 6. K Image showing the size of the tumor xenografts from three groups. L Tumor volume was measured on day 3, 6, 9, 12, 15, and 18, after OVCAR-8 cell injection. N = 6. ** p < 0.01 versus Control, ## p < 0.01 versus GPR137; error bar, SD. M Weights of xenografts derived from OVCAR-8 cells stably expressing GPR137 or in combination with hnRPD shRNA (shhnRPD). N = 6. ** p < 0.01 versus shRNA-Con+Control, ## p < 0.01 versus shRNA-Con+GPR137; error bar, SD. N Body weight of mice was examined on day 3, 6, 9, 12, 15, and 18, after OVCAR-8 cell injection. ** p < 0.01 versus Control, ## p < 0.01 versus GPR137; error bar, SD. O KI-67 staining and RAB8A staining for tumor tissues derived from OVCAR-8 cells stably expressing GPR137 or in combination with hnRPD shRNA (shhnRPD) on day 18 post inoculation. In Control group, tumor tissues were derived from OVCAR-8 cells infected with control lentiviruses in combination with lentiviruses carrying a scrambled shRNA sequence. Bar, 20 μm. P Statistical analysis of KI-67 index (%) from (F). ** p < 0.01 versus shRNA-Con+Control, ## p < 0.01 versus shRNA-Con+GPR137; error bar, SD. Q IHC score for RAB8A staining in (O). ** p < 0.01 versus shRNA-Con+Control, ## p < 0.01 versus shRNA-Con+GPR137. R The mRNA levels of RAB8A in formed tumor tissues derived from OVCAR-8 cells stably expressing GPR137 or in combination with hnRPD shRNA (shhnRPD). ** p < 0.01 versus shRNA-Con+Control, ## p < 0.01 versus shRNA-Con+GPR137; error bar, SD. S The protein levels of RAB8A in formed tumor tissues derived from OVCAR-8 cells stably expressing GPR137 or in combination with hnRPD shRNA (shhnRPD). In Control group, tumor tissues were derived from OVCAR-8 cells infected with control lentiviruses in combination with lentiviruses carrying a scrambled shRNA sequence (labeled as GPR137, -; shhnRPD, -). T qRT-PCR analysis of co-precipitated RAB8A mRNA by hnRPD antibody in the RIP assay in tumor tissues. IgG was used as a negative control. ** p < 0.01 versus IgG; error bar, SD. N = 3. U RNA pull-down assay in tumor tissues with RAB8A RNA or control RNA fragments. Protein abundance was normalized to a Tubulin

Article Snippet: GPR137 (bs-16270R, 1:1000) antibody and RAB8A (bs-6176R, 1:1000) antibody were acquired from Bioss Biotechnology (Beijing, China), while Flag-tag (M185-3, 1:10000) antibody was obtained from MBL Biotechnology (Beijing, China).

Techniques: Expressing, In Vivo, Derivative Assay, Stable Transfection, Control, Staining, Injection, Imaging, shRNA, Infection, Sequencing, Labeling, Quantitative RT-PCR, Negative Control, Pull Down Assay, Quantitative Proteomics

Journal: The Journal of Biological Chemistry

Article Title: Allosteric regulation of the Golgi-localized PPM1H phosphatase by Rab GTPases modulates LRRK2 substrate dephosphorylation in Parkinson’s disease

doi: 10.1016/j.jbc.2025.110679

Figure Lengend Snippet: PPM1H binds nonphosphorylated Rab8A and Rab10 but not Rab12 via Leu66. Microscale thermophoresis of mNeon PPM1H and mNeon L66R PPM1H with Rab10 Q68L ( A and D ), Rab8A Q67L ( B , E ), or Rab12 Q101L ( C and F ). Purified Rab proteins were serially diluted, and mNeon PPM1H or mNeon L66R PPM1H was added (final concentration of 100 nM). Graphs show the mean ± SD from three independent measurements, each using different protein preparations. Values are summarized in .

Article Snippet: Recombinant DNA , pET14b His Rab8A (Q67L) full length , Addgene , 186014; RRID: Addgene_1860014 , Reuse , .

Techniques: Microscale Thermophoresis, Purification, Concentration Assay

Journal: The Journal of Biological Chemistry

Article Title: Allosteric regulation of the Golgi-localized PPM1H phosphatase by Rab GTPases modulates LRRK2 substrate dephosphorylation in Parkinson’s disease

doi: 10.1016/j.jbc.2025.110679

Figure Lengend Snippet: Binding properties of Rab8A and thiophosphorylated Rab8A to PPM1H. A and B , thiophosphorylated Rab8A Q67L does not rely on L66 for PPM1H binding. C and D , thio-phosphorylated Q67L Rab8A but not nonphosphorylated Rab8A binds much less tightly to a PPM1H FLAP domain mutated PPM1H R338A. E and F , GTP-bound WT Rab8A binds more strongly than GDP-bound WT Rab8A to PPM1H. Various Rab8A or pRab8A proteins were serially diluted and incubated with 100 nM mNeon PPM1H variants as in . Graphs represent mean ± SD from three independent experiments using separate protein preparations. pRab, phosphoRab.

Article Snippet: Recombinant DNA , pET14b His Rab8A (Q67L) full length , Addgene , 186014; RRID: Addgene_1860014 , Reuse , .

Techniques: Binding Assay, Incubation

Journal: The Journal of Biological Chemistry

Article Title: Allosteric regulation of the Golgi-localized PPM1H phosphatase by Rab GTPases modulates LRRK2 substrate dephosphorylation in Parkinson’s disease

doi: 10.1016/j.jbc.2025.110679

Figure Lengend Snippet: Rab8A associates with liposome-bound PPM1H. Sucrose gradient coflotation of His-Rab8A Q67L (full length, nonprenylated) with mNeon PPM1H ( A , B ) or mNeon L66R PPM1H ( C , D ) in the presence and absence of 50 nm liposomes. The distribution of PPM1H and Rab8A across the gradient was determined by immunoblot; fractions were collected from the top . Quantification of three independent experiments is shown (±SD).

Article Snippet: Recombinant DNA , pET14b His Rab8A (Q67L) full length , Addgene , 186014; RRID: Addgene_1860014 , Reuse , .

Techniques: Liposomes, Western Blot

Journal: The Journal of Biological Chemistry

Article Title: Allosteric regulation of the Golgi-localized PPM1H phosphatase by Rab GTPases modulates LRRK2 substrate dephosphorylation in Parkinson’s disease

doi: 10.1016/j.jbc.2025.110679

Figure Lengend Snippet: PPM1H’s N-terminal residues contribute to Rab8A and Rab10 binding. Microscale thermophoresis of mNeon Δ37 PPM1H with His Rab8A Q67L ( A ) or Rab10 Q68L ( B ). C , sucrose gradient coflotation of His Rab8A Q67L with mNeon Δ37 PPM1H in the presence and absence of 50 nm diameter NTA(Ni) containing liposomes. D , quantification of three independent experiments is shown (±SD). Ni, nickel; NTA, nitrilotriacetic acid.

Article Snippet: Recombinant DNA , pET14b His Rab8A (Q67L) full length , Addgene , 186014; RRID: Addgene_1860014 , Reuse , .

Techniques: Binding Assay, Microscale Thermophoresis, Liposomes

Journal: The Journal of Biological Chemistry

Article Title: Allosteric regulation of the Golgi-localized PPM1H phosphatase by Rab GTPases modulates LRRK2 substrate dephosphorylation in Parkinson’s disease

doi: 10.1016/j.jbc.2025.110679

Figure Lengend Snippet: Rab8A, but not Rab12, inhibits PPM1H activity. A , anti-pRab10 immunoblot analysis of dephosphorylation by PPM1H ( upper gel ) or L66R PPM1H ( lower gel ). B and C , data were plotted as PPM1H activity, with maximum activity equal to the amount of pRab10 dephosphorylation seen in the absence of non-pRab inhibitor (lanes 3 and 4 of each gel) compared with reactions lacking PPM1H (lanes 1 and 2). D , rate of pRab10 phosphatase activity (monitored at t = 0, 8, 15, and 30 min) for mNeon-PPM1H, mNeon-L66R PPM1H, and mNeon Δ37 PPM1H. E , quantification of pRab10 intensity from ( D ), with maximum intensity detected at 0 min normalized to a value of 1. Error bars represent SD from three independent experiments; representative gels are shown. Black line , mNeon L66R PPM1H activity; red line , mNeon PPM1H activity; and blue line , mNeon Δ37 PPM1H activity. pRab, phosphoRab.

Article Snippet: Recombinant DNA , pET14b His Rab8A (Q67L) full length , Addgene , 186014; RRID: Addgene_1860014 , Reuse , .

Techniques: Activity Assay, Western Blot, De-Phosphorylation Assay